CHIMERIC ANTIGEN RECEPTOR
The present invention provides a chimeric antigen receptor (CAR) which binds a target antigen having a bulky extra-cellular domain, wherein the CAR comprises a Fab antigen binding domain. The present invention also provides nucleic acid sequences and constructs encoding such a CAR, cells expressing such a CAR and their therapeutic uses.
1 - 25 . (canceled)
26 . A method for killing a CD22-expressing target cell in a subject, the method comprising the step of administering to the subject a cell which expresses a chimeric antigen receptor (CAR) comprising an anti-CD22 antigen-binding domain which comprises:
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID No. 93)
CDR1 - NFAMA
(SEQ ID No. 94)
CDR2 - SISTGGGNTYYRDSVKG
(SEQ ID No. 95)
CDR3 - QRNYYDGSYDYEGYTMDA;
and
b) a light chain variable region (VL) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID NO: 96)
CDR1 - RSSQDIGNYLT,
(SEQ ID NO: 97)
CDR2 - GAIKLED,
and
(SEQ ID NO: 98)
CDR3 - LQSIQYP.
27 . The method according to claim 26 , wherein the antigen-binding domain of the CAR comprises a VH domain having the sequence shown as SEQ ID NO: 65; and a VL domain having the sequence shown as SEQ ID NO: 66.
28 . The method according to claim 26 , wherein the cell further comprises a second CAR which is an anti-CD19 CAR.
29 . The method according to claim 28 , wherein the antigen binding domain of the anti-CD19 CAR comprises
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID NO: 69)
CDR1 - GYAFSSS,
(SEQ ID NO: 70)
CDR2 - YPGDED,
and
(SEQ ID NO: 71)
CDR3 - SLLYGDYLDY,
and
b) a light chain variable region (VL) having CDRs with the following sequences:
(SEQ ID NO: 72)
CDR1 - SASSSVSYMH,
(SEQ ID NO: 73)
CDR2 - DTSKLAS,
and
(SEQ ID NO: 74)
CDR3 - QQWNINPLT.
30 . The method according to claim 29 , wherein the antigen-binding domain of the anti-CD19 CAR comprises a VH domain as shown in SEQ ID NO: 75 and a VL domain as shown as SEQ ID NO: 76.
31 . A method for making a cell, which comprises the step of introducing into a cell a nucleic acid sequence encoding an anti-CD22 chimeric antigen receptor (CAR) which comprises an antigen-binding domain which comprises:
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID No. 93)
CDR1 - NFAMA
(SEQ ID No. 94)
CDR2 - SISTGGGNTYYRDSVKG
(SEQ ID No. 95)
CDR3 - QRNYYDGSYDYEGYTMDA;
and
b) a light chain variable region (VL) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID No. 96)
CDR1 - RSSQDIGNYLT
(SEQ ID No. 97)
CDR2 - GAIKLED
(SEQ ID No. 98)
CDR3 - LQSIQYP
32 . The method according to claim 31 , wherein the antigen-binding domain of the CAR comprises a VH domain having the sequence shown as SEQ ID NO: 65; and a VL domain having the sequence shown as SEQ ID NO: 66.
33 . The method according to claim 31 , wherein the nucleic acid sequence comprises a first nucleic acid sequence encoding an anti-CD22 CAR and a second nucleic acid sequence encoding an anti-CD19 CAR.
34 . The method according to claim 33 , wherein the antigen binding domain of the anti-CD19 CAR comprises:
a) a heavy chain variable region (VH) having complementarity determining regions (CDRs) with the following sequences:
(SEQ ID NO: 69)
CDR1 - GYAFSSS,
(SEQ ID NO: 70)
CDR2 - YPGDED,
and
(SEQ ID NO: 71)
CDR3 - SLLYGDYLDY,
and
b) a light chain variable region (VL) having CDRs with the following sequences:
(SEQ ID NO: 72)
CDR1 - SASSSVSYMH,
(SEQ ID NO: 73)
CDR2 - DTSKLAS,
and
(SEQ ID NO: 74)
CDR3 - QQWNINPLT.
35 . The method according to claim 34 , wherein the antigen-binding domain of the anti-CD19 CAR comprises a VH domain as shown in SEQ ID NO: 75 and a VL domain as shown as SEQ ID NO: 76.
36 . The method according to claim 33 , wherein the anti-CD22 CAR is in an scFv format, the nucleic acid sequence having the general structure:
AgBD1-spacer1-TM1-endo1-coexpr-AgBD2-spacer2-TM2-endo2 or
AgBD2-spacer2-TM2-endo2-coexpr-AgBD1-spacer1-TM1-endo1,
wherein:
AgBD1 is a nucleic acid sequence encoding an antigen binding domain of the first CAR,
Spacer1 is a nucleic acid sequence encoding a spacer of the first CAR,
TM1 is a nucleic acid sequence encoding a transmembrane domain of the first CAR,
Endo1 is a nucleic acid sequence encoding an endodomain of the first CAR,
Coexpr is a nucleic acid sequence enabling co-expression of the first and second CARS,
AgBD2 is a nucleic acid sequence encoding an antigen binding domain of the second CAR,
Spacer2 is a nucleic acid sequence encoding a spacer of the second CAR,
TM2 is a nucleic acid sequence encoding a transmembrane domain of the second CAR, and
Endo2 is a nucleic acid sequence encoding an endodomain of the second CAR.